COMMON LIGHT CHAINS AND METHODS OF USE

• US 20170253670A1
• Filed: 05/19/2017
• Published: 09/07/2017
• Est. Priority Date: 11/20/2014
• Status: Abandoned Application

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Klein, Christian, Moessner, Ekkehard, Hosse, Ralf, Bruenker, Peter, Umana, Pablo, Neumann, Christiane
• Application Number:
  US15/600,015
• Publication Number:
  US 20170253670A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 39/001102 Receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: A61K 39/00117 Mucins, e.g. MUC-1
• CPC Class Codes: A61K 39/385 Haptens or antigens, bound to carriers
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 16/464 Igs containing CDR-residues from one specie grafted between FR-residues from another
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C12N 15/73 : Expression systems using phage (lambda) regulatory sequences

Abstract

The present invention generally relates to novel bispecific antigen binding molecules for T cell activation and re-direction to specific target cells comprising a common light chain. In addition, the present invention relates to polynucleotides encoding such bispecific antigen binding molecules, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the bispecific antigen binding molecules of the invention, and to methods of using these bispecific antigen binding molecules in the treatment of disease.

Claims

1. A T cell activating bispecific antigen binding molecule comprising a first and a second antigen binding moiety, wherein the first antigen binding moiety comprises a first light chain and wherein the first antigen binding moiety is capable of specific binding to an activating T cell antigen, wherein the second antigen binding moiety comprises a second light chain and wherein the second antigen binding moiety is capable of specific binding to a target cell antigen, wherein the amino acid sequence of the first and the second light chain is identical.

2. The T cell activating bispecific antigen binding molecule of claim 1, wherein the first antigen binding moiety is a Fab.

3. The T cell activating bispecific antigen binding molecule of claim 1 or 2, wherein the second antigen binding moiety is a Fab.

4. The T cell activating bispecific antigen binding molecule of any one of claims 1 to 3, further comprising a third antigen binding moiety capable of specific binding to a target cell antigen.

5. The T cell activating bispecific antigen binding molecule of claim 1, further comprising an Fc domain composed of a first and a second subunit capable of stable association.

6. The T cell activating bispecific antigen binding molecule of any one of claims 1 to 5, wherein the first and the second light chain comprises the light chain CDRs of SEQ ID NO:

32, SEQ ID NO;

33 and SEQ ID NO;

34.

7. The T cell activating bispecific antigen binding molecule of any one of claims 1 to 6, wherein the first and the second light chain comprise a kappa constant light chain domain.

8. The T cell activating bispecific antigen binding molecule of any one of claims 1 to 6, wherein the first and second light chain comprise a lambda constant light chain domain.

9. The T cell activating bispecific antigen binding molecule of claim 8, wherein the first and second light chain is a human or humanized lambda light chain.

10. The T cell activating bispecific antigen binding molecule of claim 9, wherein the first and second light chain is a human or humanized lambda 7 light chain.

11. The T cell activating bispecific antigen binding molecule of any of claims 1 to 6 and 8 to 10, wherein the first and second light chain comprise the amino acid sequence of SEQ ID NO:

31.

12. The T cell activating bispecific antigen binding molecule of claim 11, wherein the first and second light chain comprise the amino acid sequence of SEQ ID NO:

35.

13. The T cell activating bispecific antigen binding molecule of any of claims 1 to 8, wherein said antigen binding moiety capable of specific binding to an activating T cell antigen comprises a heavy chain comprising the heavy chain CDRs of SEQ ID NO:

37, SEQ ID NO;

38 and SEQ ID NO;

39.

14. The T cell activating bispecific antigen binding molecule of any of claims 1 to 13, comprising not more than one antigen binding moiety capable of specific binding to an activating T cell antigen.

15. The T cell activating bispecific antigen binding molecule of any of claims 1 to 13, wherein the first and the second antigen binding moiety are fused to each other, optionally via a peptide linker.

16. The T cell activating bispecific antigen binding molecule of any one of claims 2 to 13, wherein the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding moiety.

17. The T cell activating bispecific antigen binding molecule of any one of claims 2 to 13, wherein the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding moiety.

18. The T cell activating bispecific antigen binding molecule of any one of claims 2 to 13 and 9, wherein the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or the second subunit of the Fc domain.

19. The T cell activating bispecific antigen binding molecule of any one of claims 2 to 13, wherein the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second subunit of the Fc domain.

20. The T cell activating bispecific antigen binding molecule of any one of claims 2 to 13, wherein the first and the second antigen binding moiety are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain.

21. The T cell activating bispecific antigen binding molecule of claim 4, wherein the first, second and third antigen binding moiety are each a Fab molecule comprising an identical VLCL light chain.

22. The T cell activating bispecific antigen binding molecule of claim 4 or 21, wherein the first, second and third antigen binding moiety are each a Fab molecule comprising the light chain CDRs of SEQ ID NO:

32, SEQ ID NO;

33 and SEQ ID NO;

34.

23. The T cell activating bispecific antigen binding molecule of claim 4 or 21, wherein the first, second and third antigen binding moiety are each a Fab molecule comprising a light chain comprising the amino acid sequence of SEQ ID NO:

31.

24. The T cell activating bispecific antigen binding molecule of any one of claims 4 or 21 to 23, wherein the third antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first or second subunit of the Fc domain.

25. The T cell activating bispecific antigen binding molecule of any one of claims 4 or 21 to 23, wherein the second and the third antigen binding moiety are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, and the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding moiety.

26. The T cell activating bispecific antigen binding molecule of claim 25, wherein the second and the third antigen binding moiety and the Fc domain are part of an immunoglobulin molecule, particularly an IgG class immunoglobulin.

27. The T cell activating bispecific antigen binding molecule of any one of claims 4 or 21 to 23, wherein the first and the third antigen binding moiety are each fused at the C-terminus of the Fab heavy chain to the N-terminus of one of the subunits of the Fc domain, and the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding moiety.

28. The T cell activating bispecific antigen binding molecule of claim 27, wherein the first and the third antigen binding moiety and the Fc domain are part of an immunoglobulin molecule, particularly an IgG class immunoglobulin.

29. The T cell activating bispecific antigen binding molecule of any one of claims 5 to 28, wherein the Fc domain is an IgG, specifically an IgG₁ or IgG₄, Fc domain.

30. The T cell activating bispecific antigen binding molecule of any one of claims 5 to 25, wherein the Fc domain is a human Fc domain.

31. The T cell activating bispecific antigen binding molecule of any one of claims 5 to 23, wherein the Fc domain comprises a modification promoting the association of the first and the second subunit of the Fc domain.

32. The T cell activating bispecific antigen binding molecule of claim 31, wherein in the CH3 domain of the first subunit of the Fc domain an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the CH3 domain of the first subunit which is positionable in a cavity within the CH3 domain of the second subunit, and in the CH3 domain of the second subunit of the Fc domain an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the CH3 domain of the second subunit within which the protuberance within the CH3 domain of the first subunit is positionable.

33. The T cell activating bispecific antigen binding molecule of any one of claims 5 to 32, wherein the Fc domain exhibits reduced binding affinity to an Fc receptor and/or reduced effector function, as compared to a native IgG1 Fc domain.

34. The T cell activating bispecific antigen binding molecule of any one of claims 5 to 32, wherein the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor and/or effector function.

35. The T cell activating bispecific antigen binding molecule of claim 34, wherein said one or more amino acid substitution is at one or more position selected from the group of L234, L235, and P329.

36. The T cell activating bispecific antigen binding molecule of claim 34, wherein each subunit of the Fc domain comprises three amino acid substitutions that reduce binding to an activating Fc receptor and/or effector function wherein said amino acid substitutions are L234A, L235A and P329G.

37. The T cell activating bispecific antigen binding molecule of any one of claims 33 to 36, wherein the Fc receptor is an Fcγ

receptor.

38. The T cell activating bispecific antigen binding molecule of any one of claims 33 to 37, wherein the effector function is antibody-dependent cell-mediated cytotoxicity (ADCC).

39. The T cell activating bispecific antigen binding molecule of any one of the preceding claims, wherein the activating T cell antigen is CD3.

40. The T cell activating bispecific antigen binding molecule of any one of the preceding claims, wherein the target cell antigen is selected from the group consisting of:

Folate Receptor 1 (FolR1), Mucin-1 (MUC1), and B Cell Maturation Antigen (BCMA).

41. The T cell activating bispecific antigen binding molecule of any one of the preceding claims, wherein the first and the second light chain is a humanized light chain.

42. The T cell activating bispecific antigen binding molecule of any one of the preceding claims, wherein the first and the second light chain are capable of use in more than two antigen binding moieties having different antigen specificity.

43. The T cell activating bispecific antigen binding molecule of any of the preceding claims, wherein the target cell antigen is not BCMA.

60. An isolated polynucleotide encoding the T cell activating bispecific antigen binding molecule of any one of claims 1 to 41 or a fragment thereof or the isolated polypeptide of claim 58 or 59 or a fragment thereof.

61. A polypeptide encoded by the isolated polynucleotide of claim 60.

62. A vector, particularly an expression vector, comprising the isolated polynucleotide of claim 60.

63. A host cell comprising the isolated polynucleotide of claim 60 or the vector of claim 62.

64. A method of producing the T cell activating bispecific antigen binding molecule of any one of claims 1 to 43, comprising the steps of a) culturing the host cell of claim 63 under conditions suitable for the expression of the T cell activating bispecific antigen binding molecule and b) recovering the T cell activating bispecific antigen binding molecule.

65. A T cell activating bispecific antigen binding molecule produced by the method of claim 64.

66. A pharmaceutical composition comprising the T cell activating bispecific antigen binding molecule of any one of claims 1 to 43 and a pharmaceutically acceptable carrier.

67. A method of treating a disease in an individual, comprising administering to said individual a therapeutically effective amount of a composition comprising the T cell activating bispecific antigen binding molecule of any one of claims 1 to 43 or the light chain of any one of claims 44 to 57 in a pharmaceutically acceptable form.

68. The method of claim 67, wherein said disease is cancer.

69. A method for inducing lysis of a target cell, comprising contacting a target cell with the T cell activating bispecific antigen binding molecule of any one of claims 1 to 43 in the presence of a T cell.

44. A light chain comprising the amino acid sequence of SEQ ID NO:

32, SEQ ID NO;

33 and SEQ ID NO;

34 for use in a T cell activating bispecific antigen binding molecule.

45. The light chain of claim 44 which comprises a kappa constant light chain domain.

46. The light chain of claim 44 which comprises a lambda constant light chain domain.

47. The light chain of claim 46 which is a human or humanized lambda light chain.

48. The light chain of claim 46 which is a human or humanized lambda 7 light chain.

49. The light chain of any one of claims 46 to 48 comprising the amino acid sequence of SEQ ID NO:

31.

50. The light chain of claim 49 comprising the amino acid sequence of SEQ ID NO:

35.

51. A light chain comprising the amino acid sequence of SEQ ID NO:

32, SEQ ID NO;

33 and SEQ ID NO;

34 for use in a library for production of T cell activating bispecific antigen binding molecule.

52. The light chain of claim 51 which comprises a kappa constant light chain domain.

53. The light chain of claim 51 which comprises a lambda constant light chain domain.

54. The light chain of claim 53 which is a human or humanized lambda light chain.

55. The light chain of claim 54 which is a human or humanized lambda 7 light chain.

56. The light chain of any one of claims 53 to 55 comprising the amino acid sequence of SEQ ID NO:

31.

57. The light chain of claim 56 comprising the amino acid sequence of SEQ ID NO:

35.

58. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO:

31.

59. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO:

35.

70. A method for identifying a variable heavy chain for use in a bispecific antigen binding molecule specific for a T cell activation antigen and a target cell antigen, comprising the step of screening a combinatorial library comprising variable heavy chains with a light chain comprising the amino acid sequence of SEQ ID NO:

32, SEQ ID NO;

33 and SEQ ID NO;

34.

71. The method of claim 70, wherein the light chain comprises a kappa constant light chain domain.

72. The method of claim 70, wherein the light chain comprises a lambda constant light chain domain.

73. The method of claim 72, wherein the light chain is a human or humanized lambda light chain.

74. The light chain of claim 73 which is a human or humanized lambda 7 light chain.

75. The method of any one of claims 72 to 74, wherein the light chain comprises the amino acid sequence of SEQ ID NO:

31.

76. The method of claim 75, wherein the light chain comprises the amino acid sequence of SEQ ID NO:

35.